蓬子菜总黄酮诱导NB4细胞凋亡作用及机制  

Pro-apoptotic effect of FGVL on NB4 cells and its mechanism

作　　者：朱玉良[1] 马英丽[2] 李瑞[1] 张紫阳[1] 董均明 马玲[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院,黑龙江哈尔滨150081 [2]黑龙江中医药大学药学院

出　　处：《中国公共卫生》2017年第2期211-213,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(81274034)

摘　　要：目的观察蓬子菜总黄酮(FGVL)诱导急性早幼粒细胞白血病细胞株(NB4)细胞凋亡作用,并探讨其作用机制。方法 FGVL(50、100、200μg/mL)作用于体外培养的NB4细胞,Hoechst染色观察细胞形态学变化;流式细胞术检测细胞周期,Annexin V-FITC/PI双标记法检测细胞凋亡状况;RT-PCR检测NB4凋亡相关基因Mel-18、Sall4、Bmi-1 m RNA表达。结果细胞形态学观察显示,NB4细胞经FGVL处理48 h后,与对照组比较,FGVL各剂量组NB4细胞凋亡比例均明显升高(P<0.05),呈现细胞核固缩、核碎裂及凋亡小体等典型的细胞凋亡特征;与对照组(2.3%)比较,50、100、200μg/mL FGVL组NB4细胞凋亡率(分别为9.3%、13%、20.4%)明显升高(P<0.05);细胞周期检测结果显示,与对照组比较,50、100、200μg/mL FGVL组NB4细胞G0/G1期细胞比例(分别为43.92%、37.26%、29.15%)减少,S期细胞比例(54.16%、62.23%、70.46%)增多;与对照组比较,100、200μg/mL FGVL组NB4细胞M el-18 m RNA表达水平升高,Sall4、Bmi-1 m RNA表达水平明显降低。结论 FGVL可阻滞NB4细胞于S期,抑制细胞增殖,通过调节M el-18、Sall4、Bmi-1等凋亡相关基因表达诱导细胞凋亡。Objective To observe pro-apoptotic effect of flavone from Galium verum L （FGVL） on acute promyelocytic leukemia cells （NB4） and to explore the mechanism of the effect. Methods NB4 cells in culture medium in vitro were treated with different concentrations of FGVL （50,100, and 200 μg/mL）. Changes in cell morphological were observed with Hoechst staining and the changes in cell cycle and apoptosis rate were analyzed with flow cytometry. The expression levels of Mel-18, SaU4, and Bmi-lmRNA were assessed with reverse transcriptase PCR（ RT-PCR）. Results Morphological observations revealed that the percentages of apoptotic cells were significantly enhanced in NB4 cells treated with FGVL for48 hours,with the percentages of 9. 3% ,13% ,and 20. 4% （P 〈0.05 for all） for the dosages of 50,100, and 200 μg/mL groups, compared with that of in the control group （ 2. 3 % ） ; typical apoptotic morphological changes （karyopyknosis,karyorrhexis, and apoptosis bodies ） were observed in the NB4 cells. Cell cycle analysis indicated that compared with the control group,the cell ratios in G0/G1 phase （43.92% ,37.26% ,ans 29. 15% ） decreased significantly, while the ratios in S phase （54. 16% ,62. 23 %, and 70. 46 % ） increased obviously for the NB4 cells treated with 50,100, and 200 μg/mL FGVL. Compared with the control group, the expression level of Mel-18 mRNA increased and Sail4 and Bmi-lmRNA decreased obviously for the NB4 cells treated with 100 and 200 μg/mL FGVL. Conclusion FGVL could inhibit cell proliferation,induce cell apoptosis, and arrest cells in S phase in NB4 cells ; the mechanism of the effects is probably through regulating the expressions of Mel-18 ,Sail4 ,and Bmi-1 gene.

关 键 词：蓬子菜总黄酮(FGVL) NB4 Mel-18 SALL4 BMI-1 凋亡 

分 类 号：R557.3[医药卫生—血液循环系统疾病]